Single-Cell 5 μm-Resolution Dual-Polarity MALDI-MS Imaging without Matrix Reapplication

作者：Yanyan Chen, Rui Shi, Jianing Wang, Chengyi Xie, Yuanyuan Song, Ruxin Li, Luyao Wen, Thomas Ka Yam Lam, Zhu Yang, Zongwei Cai · 发表于：Analytical Chemistry · 年份：2025 · DOI：10.1021/acs.analchem.5c03289 · 被引用次数：10 · 研究领域：Mass Spectrometry Techniques and Applications、Advanced Proteomics Techniques and Applications、Metabolomics and Mass Spectrometry Studies

High Resolution Image Download MS PowerPoint Slide High-resolution mass spectrometry imaging (MSI) plays a vital role in lipidomics, yet challenges persist in analyzing lipids at the single-cell level due to limitations in spatial resolution and lipid coverage. While existing strategies based on a single matrix application step for dual-polarity provide high lipid coverage from the same sample and enable easy sample preparation, matrix depletion limits their spatial resolution to 10 μm, preventing their application to single-cell imaging. Here, we present a single-cell/subcellular resolution strategy for dual-polarity matrix-assisted laser desorption and ionization mass spectrometry imaging (MALDI-MSI) that eliminates the need for matrix reapplication. This approach achieves 5 μm spatial resolution while maintaining lipid coverage comparable to multistep single-cell imaging methods. This is enabled by a fine-tuned matrix deposition technique that fully utilizes the high sensitivity of N -(1-naphthyl)-ethylenediamine dihydrochloride (NEDC) in dual polarities and optimized acquisition conditions, allowing single-deposition workflows without the need for washing, repreparation, or image recalibration. This single-cell resolution MALDI-MSI strategy successfully imaged a broader range of lipid species with distinctive spatial detail in mouse kidney tissue and lung carcinoma cells (A549). Using spatial probabilistic latent semantic analysis (PLSA), we identified three distinct lipi...